- Antigen retrieval buffer
- Hydrogen peroxidase block
- High effect immunoblock
- Rabbit probe HRP
- DAB chromogen (20X DAB buffer)
- Counter strain Hematoxylin
- Antibody dilution buffer
| KIT CONTENTS | Description | Format | Recommend time |
|---|---|---|---|
| Hydrogen Peroxide Block | 3% hydrogen peroxide solution with less than 0.1% Ready to Use sodium azide. | Ready to Use | 10-20 minutes. |
| Immunoblock | PBS solution, pH 7.6, with 0.5% BSA, and less than 0.1% Ready to Use sodium azide. | Ready to Use | 10-30 minutes. |
| Rabbit Probe HRP | Conjugated HRP Reactive with rabbit primary Ready to Use antibodies. | Ready to Use | 30-45 minutes. |
| DAB Chromogen (20x) | 3,3' Diaminobenzidine (DAB) chromogen | Concentrate | Diluted with DAB Buffer (chromogen 1part: buffer 19 parts) |
| DAB Buffer | Substrate buffer, pH 7.5 with hydrogen peroxide | Ready to Use | 1-10 minutes. |
| Hematoxylin | Hydrogen peroxide | Ready to Use | 0.5-1 minutes. |
| Antigen retrieval buffer | 20X Citrate Buffer (pH6) | 20X | |
| Antibody dilution buffer | Primary antibody and incubate according to manufacturer's protocol | Ready to Use | |
Staining Protocol :
1. Deparaffinize and rehydrate formalin-fixed paraffin-embedded tissue section.
2. Add enough drops of Hydrogen Peroxide Block to cover the sections. Incubate for 10 minutes. Wash 2 times in PBS buffer.
3. Perform appropriate pretreatment if required. Wash 3 times in PBS buffer.
4. Apply Immunoblock and incubate for 10-30 minutes at room temperature to block non-specific background staining. Wash 2 times in PBS buffer.
5. Apply rabbit primary antibody and incubate according to manufacturer's protocol.
6. Wash 3 times in PBS buffer.
7. Apply Rabbit Probe HRP and incubate for 30 minutes at room temperature. Wash 3 times in PBS buffer.
8. Add 50 μl DAB Chromogen to 950ul of DAB Buffer (20X dilution), mix by swirling and apply to tissue. Incubate for 1-10 minutes. Rinse 4 times in PBS buffer.
9. Rinse slide in tap water.
10. Apply Hematoxylin counterstain according to manufacturer’s instructions (optional).
11. Dehydrate if required and cover slips.